The role of Sildenafil and Bacillus clausii for the Prevention of Methotrexate Induced Mucositis: A Preclinical Study

Abstract

Objective: Methotrexate (MTX) is a commonly used chemotherapeutic agent in clinical oncology practice and in treatment of autoimmune diseases. The major side effect during treatment is mucositis. Recently published studies reported that this effect may be due to direct intestinal epithelial injury and disturbance of intestinal microbiota. In this study, we aimed to characterize the intestinal histological changes caused by MTX and to investigate the effects and safety of Sildenafil and probiotic Bacillus clausii on intestinal mucositis in a rat model. Materials and Methods: The rats were divided in 4 groups; Group1(n=8) received MTX + B. clausii, Group 2 (n=8) received only MTX, Group3 (n=8) received MTX + Sildenafil and Group 4 (n=7) was served as control (received no treatment). Results: The rate of villus atrophy, villous fusion and erosion in the MTX + B. clausii group were found to be statistically significantly lower than in the MTX group and the MTX + Sildenafil group. Conclusion: The study demonstrated the effect of B. clausii on repairing of intestinal mucosa after MTX-treatment. The better understanding of the effects of probiotics on intestinal mucosal protection may lead to new therapeutic approaches in chemotherapy-induced mucositis.

Keywords

• Bacillus clausii
• methotrexate
• probiotic
• rat
• sildenafil

Sildenafil ve Bacillus Clausii’nin Metotreksat Bağlı Mukozitin Önlenmesindeki Rolü: Preklinik Çalışma

Öz

Amaç: Metotreksat (MTX), klinik onkoloji pratiğinde ve otoimmün hastalıkların tedavisinde yaygın olarak kullanılan bir kemoterapötik ajanıdır. Tedavi sırasındaki ana yan etkisi mukozittir. Son zamanlarda yayınlanan çalışmalar, bu etkinin doğrudan bağırsak epiteli hasarına ve bağırsak mikrobiyotasının bozulmasına bağlı olabileceğini bildirmiştir. Bu çalışmada, MTX'ın neden olduğu bağırsak histolojik değişikliklerini karakterize etmeyi ve bir sıçan modelinde Sildenafil’ in ve probiyotik Bacillus clausii’ nin bağırsak mukozit üzerindeki etkilerini ve güvenliğini araştırmayı hedefledik. Materyal ve Metot: Sıçanlar 4 gruba ayrıldı; Grup 1 (n=8) MTX + B. clausii aldı, Grup 2 (n=8) sadece MTX aldı, Grup 3 (n=8) MTX + Sildenafil aldı ve Grup 4 (n=7) kontrol olarak kullanıldı (tedavi almadı). Bulgular: MTX + B.clausii grubunda villüs atrofisi, villöz füzyon ve erozyon oranı, MTX grubu ve MTX + Sildenafil grubuna göre istatistiksel olarak anlamlı derecede düşük bulundu. Sonuç: Çalışma, MTX tedavisinden sonra B. clausii ‘nin bağırsak mukozasının onarımı üzerindeki etkisini gösterdi. Probiyotiklerin intestinal mukozal koruma üzerindeki etkilerinin daha iyi anlaşılması, kemoterapiye bağlı mukozitte yeni terapötik yaklaşımlara yol açabilir.

Anahtar Kelimeler

• Bacillus clausii
• metotreksat
• probiyotik
• sıçan
• sildenafil

References

1. Zhou B, Xia X, Wang P, et al. Induction and amelioration of methotrexate-induced gastrointestinal toxicity are related to immune response and gut microbiota. E Bio Medicine. 2018;33:122-133. doi:10.1016/j.ebiom.2018.06.029
2. Pico JL, Avila-Garavito A, Naccache P. Mucositis: its occurrence, consequences, and treatment in the oncology setting. Oncologist.1998;3(6):446-451.
3. Singh R, Zogg H, Wei L, et al. Gut microbial dysbiosis in the pathogenesis of gastrointestinal dysmotility and metabolic disorders. J Neurogastroenterol Motil. 2021;27(1):19-34. doi:10.5056/jnm20149
4. L, Chiang Chiau JS, Cheng ML, et al. SCID/NOD mice model for 5-FU induced intestinal mucositis: safety and effects of probiotics as therapy. Pediatrics and Neonatology. 2019;6(3):252-260. doi:10.1016/j.pedneo.2018.07.007
5. Yeung CY, Chiau JC, Cheng ML, et al. Immune modulation effects of lactobacillus casei variety rhamnosus on enterocytes and intestinal stem cells in a 5-FU-Induced mucositis mouse model. Gastroenterol Res Pract. 2021:3068393. doi:10.1155/2021/3068393
6. Batista VL, da Silva TF, de Jesus LCL, et al. Probiotics, prebiotics, synbiotics, and paraprobiotics as a therapeutic alternative for intestinal mucositis. Front Microbiol. 2020;11:544490. doi:10.3389/fmicb.2020.544490
7. Yeung CY, Chan WT, Jiang CB, et al. Amelioration of chemotherapy-induced intestinal mucositis by orally administered probiotics in a mouse model. PLoS One. 2015;10:e0141402. doi:10.1371/journal.pone.0138746
8. Thomsen M, Vitetta L. Adjunctive treatments for the prevention of chemotherapy- and radiotherapy-induced mucositis. Integr Cancer Ther. 2018;17:1027-1047. doi:10.1177/1534735418794885

9. Bilecik T, Ensari C, Mayir B, Cakir T, et al. Effect of sildenafil on intestinal adaptation parameters in a rat model of short bowel syndrome. Eur Rev Med Pharmacol Sci. 2016;20(3):520-527.
10. Dhayade S, Kaesler S, Sinnberg T, et al. Sildenafil potentiates a cGMP-dependent pathway to promote melanoma growth. Cell Rep. 2016;14(11):2599-2610. doi:10.1016/j.celrep.2016.02.028
11. Gautam R, Singh M, Gautam S, et al. Rutin attenuates intestinal toxicity induced by Methotrexate linked with anti-oxidative and anti-inflammatory effects. BMC Complement Altern Med. 2016;16:99. doi:10.1186/s12906-016-1069-1
12. Kolli VK, Abraham P, Rabi S. Methotrexate-induced nitrosative stress may play a critical role in small intestinal damage in the rat. Arch Toxicol. 2008;82:763-770. doi:10.1007/s00204-008-0287-9
13. Zhou B, Xia X, Wang P, et al. Induction and amelioration of methotrexate-induced gastrointestinal toxicity are related to immune response and gut microbiota. EBioMedicine. 2018;33:122-133. doi:10.1016/j.ebiom.2018.06.029
14. Chang CW, Liu CY, Lee HC, et al. Lactobacillus casei variety rhamnosus probiotic preventively attenuates 5-Fluorouracil/Oxaliplatin-induced intestinal injury in a syngeneic colorectal cancer model. Front Microbiol. 2018;9:983. doi:10.3389/fmicb.2018.00983
15. Huang L, Chiang Chiau JS, Cheng ML, et al. SCID/NOD mice model for 5-FU induced intestinal mucositis: Safety and effects of probiotics as therapy. Pediatr Neonatol. 2019;60:252-260. doi:10.1016/j.pedneo.2018.07.007
16. Cereda E, Caraccia M, Caccialanza R. Probiotics and mucositis. Current Opinion in Clinical Nutrition and Metabolic Care. 2018;21(5):399-404. doi:10.1097/MCO.0000000000000487
17. Yeung CY, Chiau JSC, Cheng ML, et al. Modulations of probiotics on gut microbiota in a 5-fluorouracil-induced mouse model of mucositis. Journal of Gastroenterology and Hepatology. 2020;35:806-814. doi:10.1111/jgh.14890
18. Urdaci MC, Bressollier P, Pinchuk I. Bacillus clausii probiotic strains: antimicrobial and immunomodulatory activities. J Clin Gastroenterol. 2004;38:S86-90. doi:10.1097/01.mcg.0000128925.06662.69
19. Lima FRF. Bacillus clausii attenuates 5-fluorouracil-induced intestinal mucositis in Mice. PhD thesis in Pharmacology - Faculty of Medicine, Federal University of Ceará, Fortaleza, Brazil.2018.
20. Picó-Monllor JA, Mingot-Ascencao JM. Search and selection of probiotics that improve mucositis symptoms in oncologic patients. A systematic review. Nutrients. 2019;11(10):2322. doi:10.3390/nu11102322